Abstract: Waste wine yeast as the research object, the superoxide dismutase (SOD) was extracted from it by toluene method.Four extraction parameters including amount of toluene, react time, react temperature and pH used for single factor test, and optimization of process parameters through L9 (34) orthogonal test.The results showed that the optimal amount of toluene were 0.8mL /g, react time was 25min, react temperature was 50℃, pH was 8.0, under these conditions, the specific activity of SOD was 178.73U /mg.The process was simple and inexpensive and coude be applied to industrialized production.